| 產品名稱 |
HP [HT1080 poly] |
| 商品貨號 |
B164628 |
| Organism |
Homo sapiens, human |
| Product Format |
frozen |
| Culture Properties |
adherent |
| Biosafety Level |
2 Cells contain CMV viral DNA sequences
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
fibrosarcoma |
| Age |
35 years |
| Gender |
male |
| Ethnicity |
Caucasian |
| Applications |
The HP polytropic retroviral packaging cell line was derived from the human fibrosarcoma cell line, HT-1080 (see ATCC CCL-121). After selection for transfected cells, individual drug resistant cell colonies were expanded, analyzed and selected for overexpression of MoMLV gag/pol. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
| Derivation |
The HP polytropic retroviral packaging cell line was derived from the human fibrosarcoma cell line, HT-1080 (see ATCC CCL-121). |
| Clinical Data |
male Caucasian 35 years |
| Comments |
The HP polytropic retroviral packaging cell line was derived from the human fibrosarcoma cell line, HT-1080 (see ATCC CCL-121). HT1080 cells were co-transfected with the methotrexate resistance vector, pFR400 and the MoMLV gag/pol expression vector, pSCV10. After selection for transfected cells, individual drug resistant cell colonies were expanded, analyzed and selected for overexpression of MoMLV gag/pol. HT1080 (clone SCV21) was co-transfected with the phleomycin resistance vector, pUT507 and the polytropic envelope expression vector, pCMVMCF and subsequently cloned. The HP clone was selected for expression of relatively high levels of both gag/pol and polytropic envelope proteins. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002.
To make the complete growth medium, add the following components to the base medium:
- O.1 mM Non-Essential Amino Acids (NEAA)
- fetal bovine serum to a final concentration of 10%
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:4 to 1:8
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| Cryopreservation |
Complete growth medium described above supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| STR Profile |
Amelogenin: X,Y CSF1PO: 12 D13S317: 12,14 D16S539: 9,12 D5S818: 11,13 D7S820: 9,10 THO1: 6 TPOX: 8 vWA: 14,19 |
| Name of Depositor |
Chiron Viagene, Inc. |
| Deposited As |
human |
| U.S. Patent Number |
|
| References |
Barber JR, et al. Retroviral packaging cell lines. US Patent 5,591,624 dated Jan 7 1997
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.
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